沉默类胰岛素多肽(Ilp)基因对褐飞虱翅和卵巢发育及海藻糖代谢的影响  

Effects of silencing insulin-like peptide(Ilp)genes on the wing and ovarian development and trehalose metabolism in Nilaparvata lugens(Hemiptera:Delphacidae)

作　　者：於卫东[1,2] 刘永康 罗雨嘉 黄镇 周泰 叶林 唐斌 王世贵[1] YU Wei-Dong;LIU Yong-Kang;LUO Yu-Jia;HUANG Zhen;ZHOU Tai;YE Lin;TANG Bin;WANG Shi-Gui(College of Life and Environmental Science,Hangzhou Normal University,Hangzhou 310036,China;Zhejiang Dingyi Biotechnology Co.,Ltd,Quzhou,Zhejiang 324100,China)

机构地区：[1]杭州师范大学生命与环境科学学院,杭州310036 [2]浙江鼎益生物科技有限公司,浙江衢州324100

出　　处：《昆虫学报》2021年第4期428-438,共11页Acta Entomologica Sinica

基　　金：国家自然科学基金项目(31672081,31371996)。

摘　　要：【目的】类胰岛素多肽(insulin-like peptide,Ilp)位于胰岛素信号通路最上游,其在糖类物质调控中发挥关键作用。本研究则旨在探究Ilp在褐飞虱Nilaparvata lugens海藻糖代谢平衡的调控作用。【方法】以褐飞虱5龄若虫为实验材料,采用RNAi技术干扰Ilps的表达,观察RNAi后褐飞虱的表型以及雌成虫的卵巢发育。RNAi 48 h与72 h后,采用生化方法测定褐飞虱5龄若虫体内海藻糖、糖原和葡萄糖含量以及海藻糖酶活性变化;采用qPCR检测海藻糖酶基因(TRE1-1,TRE1-2和TRE2)和海藻糖合成酶基因(TPS1和TPS2)的表达量变化。【结果】dsRNA可有效抑制Ilps的表达,导致褐飞虱出现异常翅型,且注射ds Ilp 3以及ds Ilp1+ds Ilp2+ds Ilp3+ds Ilp4发现褐飞虱2日龄雌成虫卵巢发育不完全。分别注射ds Ilp1-4和ds Ilp1+ds Ilp2+ds Ilp3+ds Ilp4后48 h均能显著提高葡萄糖含量;分别注射ds Ilp2,ds Ilp3及ds Ilp4后48 h显著提高了糖原含量;分别注射ds Ilp 3和ds Ilp 4后48 h能够显著提高海藻糖含量,而分别抑制Ilp2和Ilp472 h后海藻糖含量显著下降,但注射ds Ilp1+ds Ilp2+ds Ilp3+ds Ilp4后48和72 h海藻糖含量都显著上升。注射ds Ilp1+ds Ilp2+ds Ilp3+ds Ilp4后72 h可溶性海藻糖酶活性均显著上升,膜结合型海藻糖酶活性则在分别注射ds Ilp 3,ds Ilp 4和ds Ilp1+ds Ilp2+ds Ilp3+ds Ilp4后72 h显著下降;分别注射ds Ilp1,ds Ilp2和ds Ilp4后TRE1-1,TRE1-2,TRE2,TPS1和TPS2的表达量显著下降。【结论】沉默Ilp基因对褐飞虱的发育以及繁殖有一定的阻碍作用,且能够提高褐飞虱体内葡萄糖与糖原的含量,下调海藻糖酶与海藻糖合成酶基因表达量,提高可溶性海藻糖酶的活性,进而调控海藻糖的代谢。【Aim】Insulin-like peptide(Ilp)is located in the upstream of insulin signaling pathway and plays a key role in regulating sugar metabolism.This study aims to explore the function of Ilp in regulating the trehalose metabolism balance in the brown planthopper,Nilaparvata lugens.【Methods】The expression of Ilp genes in the 5th instar nymphs of N.lugens was inhibited by RNAi technology,and the phenotype and ovarian development of the female adults of N.lugens were observed after RANi.The changes in the contents of trehalose,glycogen and glucose and the trehalase activity were determined by biochemical methods,and the changes in the expression levels of trehalase genes(TRE1-1,TRE1-2 and TRE2)and trehalose synthase genes(TPS1 and TPS2)in the 5th instar nymphs of N.lugens at 48 h and 72 h after RNAi were tested by qPCR.【Results】The dsRNA effectively inhibited the expression of Ilp genes and resulted in abnormal wing in N.lugens,and the injection of ds Ilp3 and ds Ilp1+ds Ilp2+ds Ilp3+ds Ilp4 caused underdeveloped ovaries in the 2-day-old female adults of N.lugens.The glucose levels were significantly increased at 48 h after injection of ds Ilp1-4 and ds Ilp1+ds Ilp2+ds Ilp3+ds Ilp4,respectively,and the glycogen levels were also significantly increased at 48 h after injection of ds Ilp2,ds Ilp3 and ds Ilp4,respectively.The trehalose content was significantly increased at 48 h after injection of ds Ilp 3 and ds Ilp 4,respectively,while the trehalose content decreased significantly at 72 h after inhibition of Ilp2 and Ilp4,respectively,and increased significantly both at 48 h and 72 h after injection of ds Ilp1+ds Ilp2+ds Ilp3+ds Ilp4.In addition,the soluble trehalase activity increased significantly at 72 h after injection of ds Ilp1+ds Ilp2+ds Ilp3+ds Ilp4,while the membrane-bound trehalase activity decreased significantly at 72 h after injection of ds Ilp3,ds Ilp4 and ds Ilp1+ds Ilp2+ds Ilp3+ds Ilp4,respectively.The expression levels of TRE1-1,TRE1-2,TRE2,TPS1 and TPS2 were significantly decreased after inject

关 键 词：褐飞虱 类胰岛素多肽 胰岛素信号通路 海藻糖代谢 RNA干扰 

分 类 号：Q965[生物学—昆虫学]